Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18

International Journal of Current Research

and Academic Review ISSN: 2347-3215 Special Issue -4 (October-2017) Journal home page: http://www.ijcrar.com

Molecular Interaction Analysis of Mutant H-Ras P21 against Phyllanthus emblica

G. Shoba*, A. S. Pavithra and S. Ananthi

Department of Biochemistry & Bioinformatics, Dr. MGR. Janaki College of arts and science, Chennai, Tamil Nadu, India

*Corresponding author

KEYWORDS ABSTRACT

Malignant melanoma, Phyllanthus emblica Linn is widely used for medicinal purpose, usually from the plant, fresh (or) dried fruits are used. There are many bioactive compounds including H-Ras p21, Molecular docking, MAP Kinase apigenin, , , , quercetin, , , isostrictiniin, methylg allate, luteolin, emblicanin A, emblicanin B, phyllaemblicin B, and are tannins in Phyllanthus emblica. P.emblica possess several biological effects such as anticancer, antifungal, antiulcerogenic, antioxidant, and antidepressant and so on. The activation of Ras/Mitogen Activated Protein (MAP) kinase in turn deactivates MEK, a member of

the MAPK signalling cascade. The GTP-bound mutant form H-Ras (Harvey-Ras)

protein are found in 30% of human tumors and caused due to point mutation at position 12, 13, 59 and/or 61 codon. Mutant forms of H-Ras protein is continuously

involved in signal transduction for cell growth and proliferation through interaction

of downstream regulated protein Raf. The analysis of mutated H-Ras p21 with nine

compounds resulted in enormous interactions with many hydrogen bonds and least binding energy.

Introduction

Emblica officinalis is a medicinal plant and medicinal properties (Krishnaveni and commonly called as Amla (or) Indian Mirunalini., 2010). It indicate a strong gooseberry belongs to Euphorbiace ae cytotoxic action of P.emblica against cancer family. Emblica officinalis gaertn cell). Amla is rich in fibre, carbohydrate, (Phyllanthus emblica linn. Amla, Indian iron and is reported as the richest sourse of gooseberry), is widely used for medicinal vitamin-c (Singh et al., 2011).The raw fruits purpose, usually from this plant, fresh, dried of P.emblica is used for a wide variety of fruits are used. The P.emblica fruits are the human ailments including cancer. P.emblica rich source of ascorbic acid (vitamin-c) possesses several biological effects such as which itself has well documented nutritional antibacterial activity, antifungal activity,

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 antioxidant and free scavenging activity, (RAS) virus homologue was the first insecticidal activity, larvicidal and oncogene to be described in human cancer mosquitocidal activity, anticancer and anti – (Der et al., 1982). In cancer, the most proliferative activity, hepato-protective commonly mutated member of the RAS activity, antimutagenic and wound healing superfamily include HRAS, KRAS, and activity, anti ulcerogenic activity, anti NRAS. The Ras protein are small (21 depressant activity, immuno-modulatory kilodalton) G-protein that are active with activity, anti-inflammatory activity, anti- bound GTP and inactive with bound GDP. diabetic and hypoglycemic activity, Although the GTP can self-hydrolyze, there hypolipidemic activity radioprotective is a class of enzyme termed GAPs (GTPase activity and many other activity activating protein) that facilitate this (Ngamkitidechakul et al., 2010). Many hydrolysis and terminate RAS activity. Two herbal and patent drugs have been decades later,the RAS pathway still remains formulated by the constituents of this plant one of the most investigated pathways in (Rai et al 2012). E.officinalis primarily human cancer (solit et al., 2006), including contains tannins, flavonoids, phenolic melanoma, and our current understanding compounds, saponins, terpenoids, ascorbic suggest that several possible mutation along acids, carbohydrates and many other this cascade lead to tumor-promoting compounds (Khan, 2009). Supplements of physiology. These protein bind GDP/GTP fresh amla fruit is very favorable to and possess intrinsic GTPase activity individual suffering from anemia (Kumar et allowing inactivation following signal al., 2012b). This herb has many bioactive transduction in the normal cellular compounds including apigenin,gallic acid, environment (Khosravi-f et al., 1994). ellagic acid, chebulini acid, quercetin, Activation of point mutations in the Ras is chebulagic acid, corilagin,isostrictiniin, one of the most frequent genetic alterations methylgallate, luteolin, emblicanin A, associated with human cancers (Spandidos, emblicanin B, phyllaemblicin B, et al., 1984). This mutation change a single punigluconin and pednculagin are tannins amino acid in the H-Ras protein. present in emblica officinalis(Kumar et al., Specifically, the mutation replaces the 2012a). amino acid glycine with the amino acid valine at position 12 (RasG12V) (oxford, G In the post-genomic era, our understanding 2003).The amino acid residues at position of the molecular biology of melanoma has 45,46,48 , 49,50,52 and 54, near the also increased dramatically. Melanoma is a previously identified effector region, differ form of skin cancer that emanates from in the Ras (Kitayama et al .,1989). The melanocytes, which are highly specialized in amino acid position which corresponds to the formation and transfer of melanin effector region on the H-Ras is 32-40 pigment. One growth factor pathway that (Ahmadian et al., 1997).The selection of this has garnered considerable attention in the effector region as a binding site will act as last few year has been the RAS-BRAF- potential site for docking studies. Ras is MAPK-ERK signaling cascade. Much of the know to induce activation of c-RAF-1 and attention surrounding this pathway in human MAP Kinase (or) extracellular signal melanoma focuses on the fact that in regulates kinase (ERK) (De vries-smits et virtually all cases, there is an alteration at al., 1992). Such signal transducing activities some level in the RAS signaling cascade are abolished by presence of mutations in (Haluska et al., 2006). The rat sarcoma the effector regionTYR32-TYR40 (Sigal et

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 al., 1986).Mutation in the effector region isostrictiniin, methylgallate, emblicanin A, affect neither guanine-nucleotide-binding phyllaemblicin B, pedunculagin are drawn nor GTPase activity, so the effector region is in ACD/Chemsketch and then converted to considered to be region that interacts with 3D structure and saved as Mdl mol format. the target effectors of the Ras protein Then it is converted to Pdb format for (Fujita-yoshigaki et al., 1995). In second, further docking process using Open Babel. experimental results of x-ray crystallographic and nuclear magnetic Protein structure preparation resonance (NMR) analyses have shown that the three-dimensional structure of the Ras Uniprot is a freely accessible database of protein changed upon GDP to GTP protein sequence and functional information exchange (Yamasaki et al ., 1989). BRAF may entires being derived from genome that promotes RAS-RAF-MEK-ERK sequencing project. pathway activation and melanoma (http://www.uniprot.org/).The protein proliferation. Docking is a process by which databank (PDB: http://www.rcsb.org/pdb/) one can predict the significant orientation of is the single worldwide archive of structural one molecule to a second when bound to data of biological macromolecules. each other to form a stable complex. It is mostly used for finding the binding between Today depositors to the pdb have varying the ligand and the protein. The information expertise in the techniques of x-ray crystal obtained from the docking technique can be structure determination, NMR, cryoelectron used to suggest the binding energy, free microscopy and theoretical modeling. energy and stability of complex. At present, docking technique is utilized to predict the ACD/Chemsketch tentative binding parameter of ligand – receptor complex beforehand, Molecular ACD/Chemsketch is a molecular modeling docking generates different possible adduct program used to create and modify images structures that are ranked and grouped of chemical structures. Also there is a together using scoring function in the software that allows molecules and software. Docking simulations predict molecular models displayed in two and three optimized docked conformer based upon dimensions, to understand the structure of total energy of the system. In spite of all chemical bonds and the nature of the potential approaches, functional groups. ligand chemistry (tautomerism and ionization), receptor flexibility (single Openbabel conformation of rigid receptor) and scoring function (differentiate true binding mode) Open babel is a chemical tool box designed still remained the challenge. [Shafia Mir et to speak the many languages of chemical al., 2017]. data. The molecular format convert tool (open babel) is used to convert this file into Materials and Methods the pdb format and is used during analysis.

Ligand structure preparation SPDBV

The 2D structure of apigenin, gallic acid, Target and template proteins have been ellagic acid, quercetin, corilagin, loaded in SPDBV—superimposed and

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 structurally aligned. The RMSD value found Structure Retrieval to be 0.90 Å and results. The three-dimensional structure of RAS was Autodock available in the PDB database. The PDB id is 2RGB and A chain . The 3D structure Auto dock is a suite of automated docking was visualized using the Rasmol Tool and tools. The software is used for modeling shown in figure 1. RAS have the specific flexible small molecule such as drug domain region. Alternates between an molecule binding to receptor proteins of inactive form bound to GDP and an active known three dimensional structures. form bound to GTP. Activated by a guanine nucleotide-exchange factor (GEF) and It uses genetic algorithms for the inactivated by a GTPase-activating protein conformational search and is a suitable (GAP). method for the docking studies. Docking Analysis of bioactive compounds The technique combines simulated present in Phyllanthus emblicaagainst annealing for conformation searching with a mutant H-Ras and H-Ras rapid grid based method of energy evalution. The bioactive compounds including Auto dock tools are used to prepare, run and apigenin, gallic acid, ellagic acid, quercetin, analyze the docking simulations. corilagin, isostrictiniin, methylgallate, emblicanin A, and pednculagin docked Auto dock is the most cited docking against mutant H-Ras. The graphical user software because it is very fast, it provides interface program ―Auto-Dock Tools‖ was high quality prediction of ligand used to prepare, run, and analyze the conformations and good corrections between docking simulations. Kollman united atom inhibition constants and experiments ones charges, solvation parameters and polar (http hydrogens were added into the PDB file for ://autodock.scripps.edu/resources/tools) the preparation of protein in docking simulation. Autodock (Goodsell et al., 1996) Pymol requires precalculated grid maps, one for each atom type present in the flexible Pymol is computer software, a molecular molecules being docked and its stores the visualization tool use in structural biology. energy arising from the interaction with Pymol can produce high-quality 3D images rigid macromolecules. The grid box size was of small molecules and biological set at 126, 126, and 126 A0 (x, y, and z) to macromolecules such as proteins. include all the amino acid residues that present in rigid macro molecules. Auto grid Results and Discussion 4 program, supplied with Autodock 4 was used to produce grid maps (Rarey et al., Sequence Retrieval 1996). The Lamarckian Genetic Algorithm (LGA) (Morris et al., 1998) was chosen The sequence of H-Ras is retrieved from search for the best conformers. The best uniprot (http://www.uniprot.org/ uniprot/) ligand-receptor structure from the docked database and sequence accession number is structure was chosen based on the lowest P01112 from the Homosapiens (Human) energy and minimal solvent accessibility of

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 the ligand (Jones et al., 1997). The Mutant H-RAS and Gallic Acid bioactivecompounds (namely apigenin, gallic acid, ellagic acid, quercetin, corilagin, Docking of mutant H-ras against gallic acid isostrictiniin, methylgallate, emblicanin A, produced four clusters of conformers using pednculagin) and mutant H-Ras binding RMSD tolerance of 2.0 Å out of 10 docking energy are shown in Figures 1(a)-9(a), final runs. Cluster Rank 1 with binding energy - conformation from auto dock is shown in 8.48 kcal/mol at fifth run has formed eight Figures 1(b)-9(b), and the hydrogen bond hydrogen bond with active binding sites of were interactions visualized using PyMol mutant H-ras shown in the figure3. Docking were shown in Figures 1(c)-9(c). conformation between the gallic acid and mutant H-ras the is shown in Figure 3(a), Mutant H-RAS and Isostrictiniin docking score is shown in Figure 3(b).

Docking of mutant H-ras against The interactions between atoms of gallic isostrictiniin produced six clusters of acid and atoms of aminoacids of mutant H- conformers using RMSD tolerance of 2.0 Å ras is shown in Figure 3(c). Hydrogen bond out of 10 docking runs. Cluster Rank 1 with distance between the donor and acceptor binding energy -11.64 kcal/mol at eighth run atoms was shown in table-4. has formed five hydrogen bond with active binding sites of mutant H-ras shown in the Mutant H-RAS and Methylgallate figure1. Docking conformation between the isostrictiniin and mutant H-ras the is shown Docking of mutant H-ras against in Figure 1(a), docking score is shown in methylgallate produced three clusters of Figure 1(b). The interactions between atoms conformers using RMSD tolerance of 2.0 Å of isostrictiniin and atoms of aminoacids of out of 10 docking runs. Cluster Rank 1 with mutant H-ras is shown in Figure 1(c). binding energy -8.76 kcal/mol at tenth run Hydrogen bond distance between the donor has formed six hydrogen bond with active and acceptor atoms was shown in table-2. binding sites of mutant H-ras shown in the figure4. Docking conformation between the Mutant H-RAS and Quercetin methylgallate and mutant H-ras the is shown in Figure 4(a), docking score is shown in Docking of mutant H-ras against quercetin Figure 4(b). The interactions between atoms produced three clusters of conformers using of methylgallate and atoms of aminoacids of RMSD tolerance of 2.0 Å out of 10 docking mutant H-ras is shown in Figure 4(c). runs. Cluster Rank 1 with binding energy - Hydrogen bond distance between the donor 9.67 kcal/mol at first run has formed six and acceptor atoms was shown in table-5. hydrogen bond with active binding sites of mutant H-ras shown in the figure2. Docking Mutant H-RAS and Emblicanin A conformation between the quercetin and mutant H-ras the is shown in Figure 2(a), Docking of mutant H-ras against emblicanin docking score is shown in Figure 2(b). The Aproduced four clusters of conformers using interactions between atoms of quercetin and RMSD tolerance of 2.0 Å out of 10 docking atoms of aminoacids of mutant H-ras is runs. Cluster Rank 1 with binding energy - shown in Figure 2(c). Hydrogen bond 11.68 kcal/mol at sixth run has formed three distance between the donor and acceptor hydrogen bond with active binding sites of atoms was shown in table-3. mutant H-ras shown in the figure5.

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 Table.1 Showing the compounds extracted from Phyllanthus emblica

S.No Compounds Molecular 2D structure 3D structure formula

C15H10O5 1. Apigenin

2. Ellagic acid C14H6O8

3. Gallic acid C7H6O5

C15H10O7

4. quercetin

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18

5. Corilagin C27H22O18

C27H22O18

6. Isostrictiniin

C8H8O5 7. Methylgallate

8. Emblicanin A C34H22O22

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18

C33H44O19

9. Phyllaemblici n B

10. Pedunculagin C34H24O22

11. chebulinic C41H32O27 acid

12. Luteolin C15H10O6

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 Table.2 Shows the docking interaction between MUTATE H-RAS and ISOSTRINIIN

MUTATE H-RAS ISOSTRICTINIIN Distance (A) Docking energy RESIDUES ATOM (Kcal / mol) SER-17 N O 3.2 LYS-16 N O 3.0 THR-35 OG1 O 3.2 ASP-33 N O 2.8 TYR-32 N O 3.2

VAL-29 O O 2.6

ASN-116 ND2 O 3.2 ASP-119 OD1 O 2.9 -11.64 ASP-119 N O 2.7 ALA-146 OG O 2.7 SER-145 N O 3.1 LYS-147 O O 3.0 ASP-33 OD2 O 3.1 ASP-119 OD1 O 2.7 LYS-147 O O 3.3 SER-17 N O 3.0

Table.3 Shows the docking interaction between MUTATE H-RAS and QUERCETIN

MUTATE H-RAS QUERCETIN Distance (A) Docking energy RESIDUES ATOM (Kcal / mol) ASP-33 OD1 O 2.8 ASP-38 OD2 O 3.2 THR-35 OG1 O 2.6 SER-17 OG O 2.6 GLU-31 O O 2.9

GLU-31 O O 2.6

GLY-15 N O 3.1 VAL-14 N O 2.7 LYS-16 NZ O 3.6 LY-S-16 NZ O 2.9 -9.67 ASP-33 N O 3.1 THR-32 OH O 3.4 GLY-13 N O 3.1 ALA-11 O O 3.4 GLY-13 N O 2.7

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 Table.4 Shows the docking interaction between MUTATE H-RAS and GALLIC ACID

MUTATE H-RAS GALLIC ACID Distance (A) Docking energy RESIDUES ATOM (Kcal / mol) GLU-31 O O 3.4 ASP-33 O O 3.5 THR-35 OG1 O 2.5 SER-17 OG O 2.7 LYS-16 N O 2.6 GLY-15 N O 3.0 VAL-14 N O 3.3 -8.48 ASP-33 N O 3.2

LYS-16 N O 2.7

LYS-16 N O 2.9 ASP-33 O O 2.5 ASP-33 O O 3.2

Table.5 Shows the docking interaction between MUTATE H-RAS and METHYLGALLTE

MUTATE H-RAS Docking energy RESIDUES ATOM METHYLGALLATE Distance (A) (Kcal / mol) ALA-11 O O 3.4 GLY-13 N O 3.1 VAL-14 N O 2.8 GLY-15 N O 2.9 -8.76 LYS-16 N O 3.3 LYS-16 NZ O 2.6 LYS-16 NZ O 2.7 ALA-18 N O 2.8 THR-35 N O 2.7

Table.6 Shows the docking interaction between MUTATE H-RAS and EMBLICANIN A

MUTATE H-RAS EMBLICANIN A Distance (A) Docking energy RESIDUES ATOM (Kcal / mol) ASN-26 ND2 O 3.3 GLN-25 NE2 O 2.7 LYS-42 NZ O 3.3 ARG-41 NE O 2.9 GLN-25 O O 3.3 GLN-43 N O 3.0 GLN-25 NE2 O 3.2 -11.68 GLN-25 NE2 O 2.9 GLN-25 NE2 O 3.2 LYS-42 NZ O 3.3 GLN-25 O O 3.0 GLN-43 N O 3.2

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 Table.7 Shows the docking interaction between MUTATE H-RAS and ELLAGIC ACID

MUTATE H-RAS ELLAGIC ACID Distance (A) Docking energy RESIDUES ATOM (Kcal / mol) ASP-30 O O 3.0 ALA-146 N O 2.8 ASP-119 OD1 O 2.3 -7.71 ASP-30 O O 3.1 ASN-116 ND2 O 2.5 SER-145 OG O 2.5 LYS-147 N O 3.2 ALA-146 N O 3.0 ASP-119 OD1 O 3.6

Table.8 Shows the docking interaction between MUTATE H-RAS and CORILAGIN

MUTATE H-RAS CORILAGIN Distance (A) Docking energy RESIDUES ATOM (Kcal / mol) THR35 OG1 O 3.5 THR35 OG1 O 2.9 GLU31 O O 2.6 VAL29 O O 3.0

ALA11 O O 2.7

LYS16 NZ O 2.7 -12.32 GLY15 N O 2.9 VAL14 N O 2.8 ALA18 N O 2.7 LYS16 NZ O 3.3 LYS16 NZ O 2.6 ALA11 O O 3.3 GLY15 N O 3.3

Table.9 Shows the docking interaction between MUTATE H-RAS and APIGENIN

MUTATE H-RAS APIGENIN Distance (A) Docking energy RESIDUES ATOM (Kcal / mol) ASP33 O O 3.6 THR35 OG1 O 2.7 LYS16 NZ O 2.7 -8.17 SER17 OG O 2.9 GLY13 N O 3.1 ASP33 O O 2.4

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 Table.10 Shows the docking interaction between MUTATE H-RAS and APIGENIN

MUTATE H-RAS PEDUNCULAGIN DISTANCE (A) DOCKING ENERGY(Kcal RESIDUE ATOM / mol) ASN-26 ND2 O 3.3 LYS-42 NZ O 3.4 LYS-42 NZ O 3.3 ARG-41 NH2 O 3.5 ARG-41 NE O 2.8 GLN-43 NE2 O 2.9 -11.68 GLN-43 O O 3.3 GLN-43 N O 3.1 GLN-43 N O 3.0 GLN-25 O O 3.2 GLN-25 O O 2.9 GLN-25 O O 3.2

Key residues of mutant H-RAS, Hydrogen bond and Docking score

COMPOUNDS KEY RESIDUES Docking energy NO OF (Kcal / mol) INTERACTION Apigenin ASP33, THR35, LYS16, SER17, GLY13. -8.17 6 Ellagic acid ASP-30, ASN-116, SER-145, LYS-147, ALA-146, -7.71 9 ASP-119. Gallic acid GLU-31, ASP-33, THR-35,SER-17, LYS-16, -8.48 12 GLY-15, VAL-14. Corilagin THR35, GLU31, VAL29, ALA11, LYS16, GLY15, -12.32 13 VAL-14, ALA-18. Isostrictiniin SER-17, LYS-16, THR, TYR-32, VAL-29, -11.64 16 ASN-116, ASP-119 ,ALA-146, SER-145, LYS-147, ASP-33. Pedunculagin ASN-26,LYS-42,ARG-41,GLN-43, -11.68 12 LYS-42. Emblicanin A ASN-26, GLN25, LYS-42, ARG-41, GLN-25, -11.68 12 GLN-43. Quercetin ASP-33, ASP-38, THR-35, SER-17, GLU-31, -9.67 15 GLY-15, VAL-14, LYS-16, THR-32,GLY-13,ALA-11,GLY-13. Methylgallate THR35, ALA18, LYS16, GLY15, VAL14, ALA11, -8.76 9 LYS-16 GLY13.

Mutant H-RAS and Isostrictiniin

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 Mutant H-RAS and Quercetin

Mutant H-RAS and Gallic Acid

Mutant H-RAS and Methylgallate

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 Mutant H-RAS and Emblicanin A

Mutant H-RAS and Ellagic Acid

Mutant H-RAS and Corilagin

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 Mutant H-RAS and Apigenin

Mutant H-RAS and Pedunculagin

Docking conformation between the hydrogen bond with active binding sites of emblicanin A and mutant H-ras the is shown mutant H-ras shown in the figure6. Docking in Figure 5(a), docking score is shown in conformation between the ellagic acid and Figure 5(b). The interactions between atoms mutant H-ras the is shown in Figure 6(a), of emblicanin A and atoms of aminoacids of docking score is shown in Figure 6(b). The mutant H-ras is shown in Figure 5(c). interactions between atoms of ellagic acid Hydrogen bond distance between the donor and atoms of aminoacids of mutant H-ras is and acceptor atoms was shown in table-6. shown in Figure 6(c). Hydrogen bond distance between the donor and acceptor Mutant H-RAS and Ellagic Acid atoms was shown in table-7.

Docking of mutant H-ras against ellagic acid Mutant H-RAS and Corilagin produced four clusters of conformers using RMSD tolerance of 2.0 Å out of 10 docking Docking of mutant H-ras against corilagin runs. Cluster Rank 1 with binding energy - produced eight clusters of conformers using 7.71 kcal/mol at eigth run has formed four RMSD tolerance of 2.0 Å out of 10 docking

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Int.J.Curr.Res.Aca.Rev.2017; Special Issue-4: 1-18 runs. Cluster Rank 1 with binding energy - interactions between atoms of pedunculagin 12.32 kcal/mol at second run has formed six and atoms of aminoacids of mutant H-ras is hydrogen bond with active binding sites of shown in Figure 9(c). Hydrogen bond mutant H-ras shown in the figure7. Docking distance between the donor and acceptor conformation between the corilagin and atoms was shown in table-10. mutant H-ras the is shown in Figure 7(a), docking score is shown in Figure 7(b). The The bioactive compounds (apigenin, gallic interactions between atoms of corilagin and acid, ellagic acid, quercetin, corilagin, atoms of aminoacids of mutant H-ras is isostrictiniin, methylgallate, emblicanin A, shown in Figure 7(c). Hydrogen bond pedunculagin) from Phyllanthus emblica distance between the donor and acceptor docked against mutant H-Ras resulted in atoms was shown in table-8. protein and ligand complex. The docked structures were analyzed and the interaction Mutant H-RAS and Apigenin were seen. Hydrogen bond interactions and the binding distance between the donors and Docking of mutant H-ras against apigenin acceptors were measure for the best produced four clusters of conformers using conformers. The binding energy is RMSD tolerance of 2.0 Å out of 10 docking correlated with the probability of affinity runs. Cluster Rank 1 with binding energy - and stable bound between ligand and its 8.17 kcal/mol at first run has formed five receptor. Binding energy values may also hydrogen bond with active binding sites of predict the bioactivity value for ligands to mutant H-ras shown in the figure8. the corresponding receptor.

Docking conformation between the apigenin From this insilico study and previously and mutant H-ras the is shown in Figure reported experimental data in literature, we 8(a), docking score is shown in Figure 8(b). conclude this compound are least binding The interactions between atoms of apigenin energy and hydrogen bond are formed for and atoms of aminoacids of mutant H-ras is this compounds (corilagin, isostrictiniin, shown in Figure 8(c). Hydrogen bond emblicanin A, pedunculagin, quercetin) distance between the donor and acceptor would be an effective lead to inhibits atoms was shown in table-9. function of mutant H-Ras p21 protein, which will in turn arrest the process of cell Mutant H-RAS and Pedunculagin growth and proliferation of the cancer cell. Two decades later, the RAS pathway still Docking of mutant H-ras against remains one of the most investigated pedunculagin produced three clusters of pathways in human cancer, including conformers using RMSD tolerance of 2.0 Å melanoma, and our current understanding out of 10 docking runs. suggest that several possible mutation along this cascade lead to tumor-promoting Cluster Rank 1 with binding energy -11.68 physiology. Activation of point mutations in kcal/mol at ninth run has formed three the Ras is one of the most frequent genetic hydrogen bond with active binding sites of alterations associated with human cancers, mutant H-ras shown in the figure9. Docking including melanoma cancer. Further, the conformation between the pedunculagin and four ligand molecule can be incorporated mutant H-ras the is shown in Figure 9(a), into the drug development phases and docking score is shown in Figure 9(b). The clinical trial.

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How to cite this article:

Shoba G., A. S. Pavithra and Ananthi S. 2017. Molecular Interaction Analysis of Mutant H-Ras P21 against Phyllanthus emblica. Int.J.Curr.Res.Aca.Rev. Special Issue-4: 1-18.

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